title: Newly established human liver cell line: a potential cell source for the bioartificial liver in the future.
abstract: The clinical use of a bioartificial liver (BAL) device strongly depends on the development of human liver cell lines. The aim of this study was to establish and assess the potential use of the stable HepG2 cell line expressing human augmenter of liver regeneration (hALR). The cDNA encoding hALR protein was inserted into pcDNA3.1 to generate pcDNA3.1/hALR, following which pcDNA3.1/hALR was transfected to HepG2 to establish a cell line that stably expressed hALR (HepG2 hALR). A total of 800 million HepG2 hALR cells were loaded into laboratory-scale BAL bioreactors and cultured for 4 days, during which time the parameters of hepatocyte-specific function and general metabolism were determined. The cell line that stably expressed human ALR was successfully established. The expression of recombinant hALR was higher in the HepG2 hALR cell line than in the HepG2 cell line based on immunofluorescence and immunoblot assays. In samples removed from the BAL bioreactor on day 4, compared to HepG2 cells, HepG2 hALR cells produced significantly more alpha-fetoprotein (127.3 %; P < 0.05) and urea (128.8 %; P < 0.05) and eliminated more glucose (135.7 %; P < 0.05); the level of human albumin was also higher (117 %) in HepG2 hALR cells, but the difference was not significant (P > 0.05). After 24 h of culture, the mean lidocaine removal rate was 65.1 and 57.3 % in culture supernatants of HepG2 hALR and HepG2 cell lines, respectively (P < 0.01). Based on these results, we conclude that HepG2 hALR cells showed liver-specific functionality when cultured inside the bioreactor and would therefore be a potential cell source for BAL.
authors.forename: Hongling
authors.initials: H
authors.lastname: Liu
authors.forename: Shaoli
authors.initials: S
authors.lastname: You
authors.forename: Yihui
authors.initials: Y
authors.lastname: Rong
authors.forename: Yichen
authors.initials: Y
authors.lastname: Wu
authors.forename: Bing
authors.initials: B
authors.lastname: Zhu
authors.forename: Zhihong
authors.initials: Z
authors.lastname: Wan
authors.forename: Wanshu
authors.initials: W
authors.lastname: Liu
authors.forename: Panyong
authors.initials: P
authors.lastname: Mao
authors.forename: Shaojie
authors.initials: S
authors.lastname: Xin
mesh_terms: D019149:Bioreactors; D018929:Cell Culture Techniques / Q000379:methods*; D018076:DNA, Complementary / Q000235:genetics*; D056945:Hep G2 Cells; D006801:Humans; D008115:Liver Regeneration / Q000235:genetics*; D019164:Liver, Artificial; D010957:Plasmids / Q000235:genetics; D011506:Proteins / Q000235:genetics*; D014162:Transfection / Q000379:methods*; D050862:Oxidoreductases Acting on Sulfur Group Donors
chemical_list: D018076:DNA, Complementary; D011506:Proteins; C088825:GFER protein, rat; D050862:Oxidoreductases Acting on Sulfur Group Donors